Modified dog alpha1-interferon gene, construction method of expression vector and preparation method of protein encoded by modified dog alpha1-interferon gene

Abstract

The invention discloses a modified dog alpha1-interferon gene and a preparation method of dog alpha1-interferon. The modified dog alpha1-interferon gene has the nucleotide sequence as shown in SEQ ID NO.1 and the amino acid sequence coded by the gene is shown in SEQ ID NO.2. The invention concretely relates to matching and modification for the predilection of a dog alpha1-interferon codon to a yeast codon as well as optimal control on a culture method for efficiently producing dog alpha1-interferon, pollution avoiding measures and operation methods of methanol adding amount and time determination, protein expression time control, namely protein harvest time, sterility test, protein content and activity measurement, culture condition expansion and the like. The method disclosed by the invention is simple and easy to realize, relatively low in cost, and capable of realizing stable and efficient expression of the dog alpha1-interferon in pichia pastoris, and the expressed dog alpha1-interferon has a remarkable effect on treating dog viral infectious diseases and can enhance the immune effect of a rabies vaccine.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals. More specifically, it relates to a modified canine α1-interferon gene, the construction of expression vector and the preparation method of its protein. Background technique [0002] Interferon (IFN) is a glycoprotein with broad-spectrum antiviral activity produced by cells under the action of a specific inducer. When it acts on other cells again, it can immediately obtain antiviral, antitumor and other aspects immunity. A large number of in vitro and in vivo tests have shown that canine α1-interferon has protective and inhibitory effects on viral infectious diseases that are a major threat to production. Interferon has broad application prospects due to its wide range of effects and small residues. However, due to the high cost of interferon and the economic value of animals, its clinical application in veterinary medicine is mainly limited to the treatment of pet diseases, and the ...

AI Technical Summary

Problems solved by technology

[0004] Although some achievements have been made in the use of yeast to express interferon, how to modify the interferon gene to make it biased from yeast for different genes and homologous genes from different species Phenotropic matching to increase the expression of interferon; how to cultivate during the expression process, how to ensure the dissolved oxygen required for protein expression, how to avoid pollution, the amount and time of methanol induction, and the control of protein expression time Harvesting time, methods of sterility testing, protein content and activity determination are quite different, and a stable and complete technical solution has not been formed, which is not conducive to the development of industrialized production of canine α1-interferon, and cannot fundamentally Solve the cost problem of canine α1-interferon, which affects its popularization and application

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